Anti-VDAC1/Porin antibody [16G9E6BC4] - Mitochondrial Loading Control

• WB

Unknown

Western blot - Anti-VDAC1/Porin antibody [16G9E6BC4] - Mitochondrial Loading Control (AB110326)

All lanes:

Western blot - Anti-VDAC1/Porin antibody [16G9E6BC4] - Mitochondrial Loading Control (ab110326) at 1 µg/mL

All lanes:

Yeast spheroplast membrane fraction.

Predicted band size: 31 kDa

false

• WB

CiteAb

Western blot - Anti-VDAC1/Porin antibody [16G9E6BC4] - Mitochondrial Loading Control (AB110326)

VDAC1/Porin western blot using anti-VDAC1/Porin antibody [16G9E6BC4] - Mitochondrial Loading Control ab110326. Publication image and figure legend from Shai, N., Yifrach, E., et al., 2018, Nat Commun, PubMed 29720625.

ab110326 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab110326 please see the product overview.

Fzo1 has characteristics of a tether for the PerMit contact site. a Overexpression of full-length (mCherry-Fzo1) or truncated (mCherry-Fzo1 Δ547) Fzo1 (control cells and schemes of Fzo1 on left). When full-length Fzo1 is overexpressed, mitochondria (marked by MTS-BFP) are hyper-fused (as previously described) yet Fzo1 is still only localized to mitochondrial patches in proximity to peroxisomes and to the contact (white arrows). All PerMits co-localize with Fzo1. Overexpression of truncated Fzo1 does not alter mitochondrial morphology, is sufficient for mediating PerMit expansion and enables Fzo1 localization to the PerMit site. Overexpression of full length or truncated Fzo1 does not affect peroxisomes (marked by CFP-SKL) number and size. b Quantification of peroxisomes that co-localize with mitochondria in control, full length or truncated Fzo1 overexpression. Bars represent the mean ± s.e. from three independent experiments. one-tail Student's t-test, **p < 0.01. c Total lysates from control and Δmdm30 cells, in which native Fzo1 is tagged with 13Myc epitopes (Fzo1-13Myc), were subjected to subcellular fractionation yielding soluble (Sup) and mitochondria-enriched (Pellet) fractions. The distinct fractions were analyzed by immunoblot with anti-Myc (Fzo1), anti-Pgk (a cytosolic marker) and anti-Porin (a mitochondrial marker) antibodies. Fzo1 is found in a non-mitochondrial fraction more readily visible when a slight increase in Fzo1 levels is created by deleting MDM30. d Fzo1 overexpression by a strong TEF2 promoter demonstrates Fzo1 signals that do not co-localize with mitochondria (marked by Tom70-GFP) but rather with peroxisomes (marked by CFP-SKL) (white arrows). eMDM30 depletion demonstrating that Fzo1-GFP is localizes to both mitochondria (marked by DAPI) and peroxisomes (marked by RFP-SKL) (white arrows). f A physical interaction between Fzo1 and Pex14/Pex19. HA-Fzo1 or its corresponding empty vector were expressed in control, GFP-Pex14, or GFP-Pex19 strains. HA-Fzo1 was immunoprecipitated from solubilized crude membrane extracts using HA-coupled beads. Eluted Fzo1 was analyzed by SDS-PAGE and immunoblot using anti HA- or GFP-specific antibodies. Ponceau S staining (PoS) was used to compare protein levels

false